Genetic Influences in Mouse Spermatogonial Stem Cell Self-Renewal
Genetic Influences in Mouse Spermatogonial Stem Cell Self-Renewal
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DOI：
10.1262/jrd.09-153n
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发表时间：
2010-02-01
影响因子：
1.8
通讯作者：
Shinohara, Takashi
中科院分区：
文献类型：
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作者：
Kanatsu-Shinohara, Mito;Ogonuki, Narumi;Shinohara, Takashi
精原干细胞是一种缓慢分裂的细胞，具有自我更新的分裂功能。虽然遗传背景在干细胞自我更新中的作用在造血干细胞中已经得到了很好的研究，但对其在其他自我更新组织（包括SSCs）中对干细胞的作用知之甚少。为了研究遗传因素是否参与SSC自我更新的调节，我们首先研究了不同近交系小鼠（C57 BL/6，DBA/2，AKR，BALB/C和C3 H）在白消安引起的化学损伤后的精子发生。DBA/2和AKR品系的精子发生对白消安治疗相对耐受，而C5713 L/6小鼠的精子发生减少，C3 H和BALB/C小鼠的精子发生几乎消失。系列生殖细胞移植实验提供了功能证据，即具有DBA/2背景的SSC比具有B6背景的SSC扩增更快。最后，我们还采用了生殖干细胞（GS）培养技术来检测体外自我更新活性。虽然GS细胞的遗传操作已被限制到那些从DBA/2背景，我们产生的C3 H背景的转基因后代的GS细胞与质粒载体的电穿孔。我们的研究结果强调了遗传因素在SSC自我更新中的重要性。此外，将遗传修饰技术应用于具有非DBA/2背景的GS细胞扩展了基于SSC的方法在雄性生殖系修饰中的潜力。
Spermatogonial stern cells (SSCs) are slowly dividing cells that undergo self-renewal division to support spermatogeriesis. Although the effects of genetic background in stem cell self-renewal have been well studied in hematopoietic stem cells, little is known about its effect on stem cells in other self-renewing tissues, including SSCs. To examine whether genetic factors are involved in regulation of SSC self-renewal, we first studied spermatogenesis in different inbred Mouse strains (C57BL/6, DBA/2, AKR, BALB/C and C3H) after chemical damage caused by busulfan. Spermatogenesis in the DBA/2 and AKR strains was relatively resistant to busulfan treatment, whereas spermatogenesis was diminished in C5713L/6 mice and nearly ablated in C3H and BALB/C mice. Serial germ cell transplantation experiments provided functional evidence that SSCs with the DBA/2 background expanded more rapidly than those with the B6 background. Finally, we also employed the Germline Stem (GS) cell culture technique to examine the self-renewal activity in vitro. Although genetic manipulation of GS cells has been limited to those from the DBA/2 background, we produced transgenic offspring of the C3H background by electroporation of GS cells with a plasmid vector. Our results underscore the importance of genetic factors in SSC self-renewal. Furthermore, application of genetic modification techniques to GS cells with non-DBA/2 backgrounds extends the potential of a SSC-based approach in male germline modification.